Effect of CIDEa Overexpression on Lipid Synthesis of Mammary Gland in Goats (Capra hircus)
SUN Yu-Ting1, LI Pei-Wei1, SU Li-Na1, HU Han1, WU Xiao-Min1, LI Jun2,*
1 Shaanxi Institute of Zoology, Xi'an 710032, China; 2 College of Animal Science and Technology, Henan University of Animal Husbandry and Economy, Zhengzhou 450046, China
Abstract Cell-death-inducing DNA fragmentation factor-α-like effector a (CIDEa) is a lipid droplet associated protein, which is involved in regulating lipid metabolism and maintaining body energy balance. In order to further explore the function of CIDEa in lipid metabolism in mammary gland of dairy goats (Capra hircus), in this study, dairy goat mammary epithelial cells were used as a model to study the effect of CIDEa gene on lipid synthesis by constructing pcDNA3.1-CIDEa overexpression vector. The results showed that the mRNA expression of CIDEa gene was increased nearly 10-fold after 48 h of transfection with pcDNA3.1-CIDEa in dairy goat mammary epithelial cells (P<0.01). The results of triacylglycerol (TAG) contents assay and Oil Red O staining showed that the accumulation of lipid droplets and the content of triglyceride (P<0.05) in cells were increased after CIDEa overexpression. The results of real-time quantitative PCR showed that CIDEa overexpression significantly increased the expression of acetyl-CoA carboxylase alpha (ACACA)(P<0.05), cluster of differentiation 36 (CD36)(P<0.05), sterol regulatory element binding protein 1 (SREBP1)(P<0.05) and diacylglycerol acyltransferase 2 (DGAT2)(P<0.01).These results indicated that CIDEa gene could promote lipid synthesis in mammary epithelial cells. This study provides a theoretical basis for further studying the role of CIDEa gene in the regulation of milk fat metabolism in dairy goats.
SUN Yu-Ting,LI Pei-Wei,SU Li-Na, et al. Effect of CIDEa Overexpression on Lipid Synthesis of Mammary Gland in Goats (Capra hircus)[J]. 农业生物技术学报, 2024, 32(3): 587-594.
[1] 李君, 孙雨婷, 郭震楠, 等. 2022. 干扰CIDEa基因对奶山羊乳腺上皮细胞脂质合成的影响[J]. 农业生物技术学报, 30(05): 918-925. (Li J, Sun Y T, Guo Z N, et al.2022. Effect of CIDEa interference on lipid synthesis of mammary epithelial cells in dairy goats (Capra hircus)[J]. Journal of Agricultural Biotechnology, 30(05): 918-925.) [2] 李君, 吴姣, 权凯, 等. 2020. 奶山羊CIDEa基因的克隆、序列分析与组织表达[J]. 畜牧与兽医, 52(03): 6-11. (Li J, Wu J, Quan K, et al.2020.Cloning, sequence analysis and tissue expression of the CIDEa gene in dairy goat[J]. Animal Husbandry & Veterinary Medicine, 52(3): 6-11.) [3] 王桢, 罗军, 王伟, 等. 2010. 奶山羊乳腺上皮细胞的分离、培养及鉴定[J]. 生物工程学报, 26(08): 1123-1127. (Wang Z, Luo J, Wang W, et al.2010. Characterization and culture of isolated primary dairy goat mammary gland epithelial cells[J]. Chinese Journal of Biotechnology, 26(8): 1123~1127) [4] 张敏. 2016. 能量负平衡奶牛肝脂沉积过程中SREBP-1c/CIDEA通路的作用机制[D]. 硕士学位论文, 吉林大学, 导师: 李小兵, pp. 27-38. (Zhang M.2016. The mechanism of SREBP-1c/Cidea pathway in the hepatic lipidosis of dairy cows with negative energy balance[D]. Thesis for M.S., Jilin University, Supervisor: Li X B, pp. 27-38.) [5] Belal S A, Kang D R, Sivakumar A S, et al.2019. Effect of long chain fatty acids on triacylglycerol accumulation, fatty acid composition and related gene expression in primary cultured bovine satellite cells[J].Animal Biotechnology, 30(4): 323-331. [6] Bionaz M, Loor J J.2008. Gene networks driving bovine milk fat synthesis during the lactation cycle[J]. BMC Genomics, 9: 366. [7] Cheng J, Xu D, Chen L, et al.2022. CIDEA regulates de novo fatty acid synthesis in bovine mammary epithelial cells by targeting the AMPK/PPARγ axis and regulating SREBP1[J]. Journal of Agricultural and Food Chemistry, 70(36): 11324-11335. [8] Chitraju C, Walther T C, Farese R V Jr.2019.The triglyceride synthesis enzymes DGAT1 and DGAT2 have distinct and overlapping functions in adipocytes[J]. Journal of Lipid Research, 60(6): 1112-1120. [9] Guo P P, Jin X, Zhang J F, et al.2023.Overexpression of DGAT2 regulates the differentiation of bovine preadipocytes[J].Animals (Basel),13(7): 1195. [10] Haenlein G F W.2001. Past, present, and future perspectives of small ruminant dairy research[J]. Journal of Dairy Science, 84(9): 2097-2115. [11] Haenlein G F W.2004. Goat milk in human nutrition[J]. Small Ruminant Research, 51: 155-163. [12] Hao J W, Wang J, Guo H L, et al.2020. CD36 facilitates fatty acid uptake by dynamic palmitoylation-regulated endocytosis[J]. Nature Communications, 11(1): 4765. [13] Inohara N, Koseki T, Chen S, et al.1998. CIDE, a novel family of cell death activators with homologyto the 45 kDa subunit of the DNA fragmentation factor[J]. The EMBO Journal, 17(9): 2526-2533. [14] Ji Z B, Chao T L, Zhang C L, et al.2019. Transcriptome analysis of dairy goat mammary gland tissues from different lactation stages[J]. DNA and Cell Biology, 38(2): 129-143. [15] Li C, Zhu J J, Shi H B, et al.2020. Comprehensive transcriptome profiling of dairy goat mammary gland identifies genes and networks crucial for lactation and fatty acid metabolism[J]. Frontiers in Genetics, 11: 878. [16] Ma L, Corl B A.2012. Transcriptional regulation of lipid synthesis in bovine mammary epithelial cells by sterol regulatory element binding protein-1[J]. Journal of Dairy Science, 95(7): 3743-3755. [17] Park Y W.1994. Hypo-allergenic and therapeutic significance of goat milk[J]. Small Ruminant Research, 14: 151-159. [18] Rudolph M C, Neville M C, Anderson S M.2007. Lipid synthesis in lactation: Diet and the fatty acid switch[J]. Journal of Mammary Gland Biology and Neoplasia, 12(4): 269-281. [19] Wang R, Kong X, Cui A, et al.2010. Sterol-regulatory-element-binding protein 1c mediates the effect of insulin on the expression of Cidea in mouse hepatocytes[J]. Biochemical Journal, 430(2): 245-254. [20] Wang W S, Lv N, Zhang S S, et al.2012. Cidea is an essential transcriptional coactivator regulating mammary gland secretion of milk lipids[J]. Nature Medicine, 18(2): 235-243. [21] Xu H F, Luo J, Zhao W S, et al.2016. Overexpression of SREBP1 (sterol regulatory element binding protein 1) promotes de novo fatty acid synthesis and triacylglycerol accumulation in goat mammary epithelial cells[J]. Journal Dairy Science, 99(1): 783-795. [22] Yonezawa T, Haga S, Kobayashi Y, et al.2009. Saturated fatty acids stimulate and insulin suppresses CIDE-A expression in bovine mammary epithelial cells[J]. Biochemical and Biophysical Research Communications, 384(4): 535-539. [23] Yonezawa T, Yonekura S, Kobayashi Y, et al.2004. Effects of long-chain fatty acids on cytosolic triacylglycerol accumulation and lipid droplet formation in primary cultured bovine mammary epithelial cells[J]. Journal of Dairy Science, 87(8): 2527-2534. [24] Zhang S S, Shui G H, Wang G Q, et al.2014. Cidea control of lipid storage and secretion in mouse and human sebaceous glands[J]. Molecular and Cellular Biology, 34: 1827-1838. [25] Zhou Z H, Yon Toh S, Chen Z M, et al.2003.Cidea-deficient mice have lean phenotype and are resistant to obesity[J]. Nature Genetics, 35(1): 49-56.
